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KMID : 0043320020250010080
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Archives of Pharmacal Research
2002 Volume.25 No. 1 p.80 ~ p.85
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Spontaneous Release of Glycosylphosphatidylinositol (GPI)- anchored Renal Dipeptidase from Porcine Renal Proximal Tubules
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Choi K
Park SW/Kang BY/Yoon HJ/Park EM/Lee HB/Hooper NM/Park HS
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Abstract
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The incubation of porcine renal proximal tubules (PTs) resulted in the release of the glycosylphosphatidylinositol (GPI)-anchored renal dipeptidase (RDPase, EC 3. 4. 13. 19) from the membrane after a lag period of approximately 6 hours. This spontaneous release of RDPase from the membrane was inhibited by antibiotics. When the incubation supernatant was added back to fresh PTs, both the antibiotic inhibition of RDPase release and the lag period disappeared . The released RDPase reacted with an anti-cross reacting determinant antibody indicating the presence of the Ins (1,2-cyc)P moiety. These results suggest that bacteria in the PTs, when incubated, grow and secrete a phosphatidylinositol-specific phospholipase C (Pl-PLC). This enzyme then hydrolyses the GPI-anchored RDPase arid is transferable. RDPase was purified following its release from the membrane by this simple and inexpensive method which may also be applied to other GPI-anchored proteins.
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